Genomic DNA sequencing of a Holstein male
Genomic DNA sequencing of a Holstein male
Blood samples were collected from jugular vein using K2-EDTA blood collection tubes. DNA was isolated from a blood sample using Quick-DNA Miniprep Plus kit (Zymo research) following manufacturer's instructions. Genomic DNA library was prepared using 50 ng of DNA (for a sample with a word, gDNA, in a file name) - 100 ng (gDNA2X) isolated from a Holstein male (HOL1_m) by following Nextera DNA Sample Preparation Guide (Illumina). The tagmented DNA was purified and eluted in 21 ul elution buffer using the MinElute PCR purification kit. The DNA was amplified using NEBNext High-Fidelity 2X PCR Master Mix with Ad1 and Ad2 primers (Buenrostro et al., 2013) for 5 PCR-cycles. The amplified libraries were purified and eluted in 51 ul elution buffer using the MinElute PCR purification kit. Library size distribution was monitored using 10 ul of the library by QIAxcel capillary electrophoresis (Qiagen). Large DNA fragments were eliminated using AMPure XP magnetic beads (Beckman Coulter, Brea, CA) by a right-side size selection using 0.55 x followed by 1.5 x ratio of beads to sample. Library concentration was estimated using KAPA Library Quantification kit (Kapa Biosystems, Wilmington, MA).